mRNA Stability and Translation Optimization

In vitro-transcribed (IVT) mRNA is designed to structurally resemble naturally occurring mature and processed mRNA in the cytoplasm, translating the message to the desired corresponding protein, the pharmacologically active product. The first pre-clinical use of mRNA was in the 1990s. Nowadays, owing to its unique advantages, the application prospect of IVT mRNA is broad, including cancer immunotherapy, infectious disease vaccines, protein replacement therapy, and cell engineering. However, its development faces two major technical obstacles. Firstly, IVT mRNA is intrinsic instability. It is easily degraded by extracellular endonuclease and thus difficult to be effectively internalized. Secondly, the translation of IVT mRNA is inefficient in the body. At present, the target of mRNA drug delivery system is mostly confined to liver, and the breakthrough still lies in the development of delivery technology. Creative Biogene is a leading customized service provider in mRNA-based drugs research and development (R&D). To deal with the challenges of IVT mRNA, we offer a series of sophisticated pharmaceutical technologies and efficient delivery systems.

Fig1. Principles of antigen-encoding mRNA pharmacology. (Sahin, U., et al, 2014)

Strategies for optimizing mRNA stability and translation in Creative Biogene

Backbone optimization

Stable translation of IVT mRNA requires a functional 5' cap structure. The structure has an impact on innate sensing and protein production. Synergy with the 5'cap and other determinants, the poly(A) tail regulates the stability and translation efficiency of the IVT mRNA through its length. In addition, other elements of IVT mRNA, including the codon composition of the region (ORF, its dinucleotides composition), and untranslated region (UTR, their sequences, length and secondary structures), also are important for the regulation of the translation of the mRNA and the protein expression. Our strategy is to work closely around the fundamental elements of IVT mRNA, involving synthetic cap analogues and capping enzymes, engineering of UTRs and ORF, elongation of the poly(A) tail, and/or incorporation of modified nucleosides. These optimization strategies can be offered as a stand-alone service or integrated to provide the best combination.

Separation and/or purification techniques

In order to increase mRNA translation, we also apply separation and/or purification techniques, including RNase III treatment and fast protein liquid chromatography (FPLC) purification. Other methods, such modulation of target cells (co-delivery of translation initiation factors) can also be conducted according to the clients' specific project needs.

mRNA intracellular delivery system

In addition to optimizing an mRNA construct, in vivo mRNA delivery is critical to increasing mRNA stability and translation. Because mRNA is a transient molecule by nature and is susceptible to degradation by nuclease activity, thus efficient protection is required. Integrating cutting-edge platforms as well as substantial experience in the study of mRNA delivery system, we have the capability to regulate mRNA stability and translation through in vitro and in vivo strategies. For in vivo strategies, we are not only able to perform studies of the current mature delivery systems (such as lipids-based delivery system, polymers-based delivery system and hybrid system), but also develop novel delivery system for the increasing needs of mRNA-based therapy. For in vitro strategies (the improved injection strategies), such as electroporation of mRNA and gene gun-based administration, are offered in our platform.

mRNA therapeutics has shown great potential as a novel class of drug modalities. With the help of Creative Biogene' highly efficient strategies and a range of in-depth services, our customers' research projects can be supported in a timely and professional manner. For more detailed information, please feel free to contact us or directly send us an inquiry.